34results about How to "Increase screening throughput" patented technology

The invention discloses a rapid screening method for comprehensively evaluating the in vitro inhibitory effects of 9 human liver CYP450 metabolic enzymes by using 14 kinds of probe substrates and 16 kinds of probe reactions. The invention mainly relates to monitoring the metabolic activity changes of 9 kinds of human liver CYP450 enzymes by using an in vitro mixed probe incubation method combined with LC / MS / MS, and rapidly and comprehensively evaluating the inhibitory effect of test compounds on metabolic enzymes. This method comprehensively considers the specificity and diversity of probe substrates, the interaction between probe substrates, and the reaction of different incubation conditions (organic solvent, buffer, BSA, etc.) in the incubation system to the probe reaction. The influence of various factors such as the influence on the activity of the enzyme, and the enzyme kinetics characteristics of the 16 probe reactions under the selected incubation conditions, combined with the high sensitivity and high selectivity LC-MS / MS technology, established a new in vitro evaluation System, in order to more accurately and comprehensively predict the possible inhibitory effects of test compounds on 9 major metabolic enzymes of human liver in the high-throughput screening of new drug development, and improve the predictability of later metabolic interactions.

The invention relates to the technical field of microorganisms, and specifically discloses a Saccharopolyspora spinosa DS190375 and its fermentation product, bacterial agent, fermentation and screening method and application. The Saccharopolyspora spinosa DS190375 of the present invention has a preservation number of CGMCC No.18681. The fermentation method is as follows: the bacteria are rejuvenated and screened in a high-salt plate culture medium, and then fermented. The present invention also provides a screening method for a class of S. spinosa with high spinosad yields. First, screen the obtained S. spinosa with high salt tolerance, and then conduct the screening on the obtained S. spinosa with high salt tolerance. Screening to obtain strains with high spinosad production. The Saccharopolyspora spinosa DS190375 of the present invention is resistant to high salt and has high spinosad yield. The fermentation method avoids the problem of unstable spinosad yield caused by strain degradation.

The invention relates to a phage antibody library which assembles an ScFv gene fragment between Restriction Enzyme cutting sites of SfiI and NotI of a pCANTAB5E carrier. The phage antibody library is characterized in that the ScFv gene fragment can be affinitive and enriched with antigens of 16-membered macrolide agricultural chemicals to form a soluble single-chain antibody and the phage antibody library is applied to immunoassay of pesticide residue. The phage antibody library has the advantages that the antibody with high affinity can be obtained without animal immunization, the test period is short, the antibody library is the phage antibody library which takes small molecular milbemycin oxime of a 16-membered macrolide generic structure as immunogen to construct, the antibody librarytheoretically can directly obtain a specific antibody library of 16-membered macrolide compound through screening, the screening flux is high, the efficiency is high, the specificity is strong, and the affinity selecting range is wide, so that the phage antibody library has wide application prospect in the aspects such as agricultural chemical antibody preparation, testing technique development and the like.

The invention provides a semi-contact under-oil continuous droplet sample applying and liquid adding method, which is suitable for sequentially operating a droplet array system. According to the method, the distance between the pointed end of a capillary tube sample applying needle and the lower surface of a micro-hole (or a generated droplet) is controlled accurately, and the affinity or interface tension interaction between the droplet and the surface (or the generated droplet) is utilized, so that rapid and reliable continuous droplet sample applying or continuous liquid adding is realized, and the problem of cross infection during sample applying is solved effectively. The method is suitable for biochemical analysis screening researches such as high-flux medicament screening, protein crystallization condition screening, enzyme kinetics research and drug toxicity determination.

The invention relates to the field of genetic resource conservation, particularly relates to the field of animal paternity identification, and discloses a method of establishing an animal paternity identification system on basis of whole genome STR. The method comprises the following steps: 1) looking up whole genome STR loci; 2) preliminarily screening the STR loci; 3) carrying out a primer design; 4) randomly sampling animal populations and extracting genome DNA; 5) verifying the population diversity of the STR loci; 6) carrying out Hardy-Weinberg equilibrium inspection on the STR loci with genetic polymorphism, and screening out the STR loci with the genotype frequency distribution which meets the Hardy-Weinberg equilibrium; 7) evaluating the forensic medicine application value of the STR loci; 8) establishing the STP animal paternity identification system. The method is convenient to operate; the constructed paternity identification system has a paternity exclusion probability of more than 0.99, and is capable of accurately carrying out animal paternity identification, therefore, the paternity identification system can be used for guiding the breeding of rare species, and the identification and breeding of economic species.

The invention provides high-yield coenzyme Q10 rhodobacter sphaeroides and mutation breeding and application thereof, and relates to the field of microbial mutation breeding. Strains screened by spacemutagenesis are used as starting bacteria and subjected to plasma mutagenesis at normal pressure and room temperature. The high-yield coenzyme Q10 rhodobacter sphaeroides are obtained after multi-resistance screening, high-throughput screening and step-by-step amplification verification, and are deposited in China General Microbiological Culture Collection Center with the deposit number of CGMCCNo. 16625. The average titer of a mutant strain of the rhodobacter sphaeroides can reach 2694[mu] / mL according to 20m<3> fermentation production verification, and the yield of the coenzyme Q10 is significantly improved. In addition, the strain has quadruple resistance and broad application prospects.

The invention is suitable for the technical field of genetic engineering, and provides a target specific influenza A virus medicament screening yeast two-hybrid system and application thereof. The target specific influenza A virus medicament screening yeast two-hybrid system comprises an influenza A virus PB1gene and an influenza A virus gene, a PB1 gene and a PA gene, and PB1, PB2 and PA genes. The target specific influenza A virus medicament screening yeast two-hybrid system for medicament screening has the advantages of high specificity, high throughput screening and exact mechanism of a target medicament.

The invention relates to an enzyme screening method, particularly to a screening method for a halohydrin dehalogenase. Halohydrin dehalogenase with high enzyme activity is screened by pH color development and a primary screening-secondary screening two-step method. By employing a deep-hole plate for microminiaturized culture, the invention realizes high throughput culture and high throughput preparation of a crude enzyme liquid. PH change is utilized as a breakthrough to screen halohydrin dehalogenase with high enzyme activity, the efficiency is high, the cost is low, and the speed is fast. Fewer media and reagents are consumed. The method provided by the invention makes mechanized automatic operation come true, and with the help of an automated high-throughput screening instrument, the amount of labor can be greatly reduced further, the screening flux can be improved.

Disclosed are a 3-furyl-2-cyano-2-acrylamide derivative with epidermal growth factor receptor (EGFR) inhibitory activity and pharmaceutical acceptable salt thereof, together with preparation method thereof, pharmaceutical composition comprising the compound, and application of the compound in treating senile dementia (AD). The new compound is shown as formula I, wherein R1 is selected from the group consisting of H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl and C3-C6 cycloalkyl; R2 is selected from the group consisting of alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl and heteroaralkyl; X is selected from the group consisting of CH2, NH, O and S; m and n are all integers greater than or equal to zero.

The invention relates to a screening method and application of microorganisms containing MVA and / or MEP metabolic pathways, belonging to the field of biosynthesis and fermentation engineering. The above-mentioned screening method comprises the following steps: inoculate the spore suspension of microorganisms into a culture container for cultivation to obtain single colonies of microorganisms, cultivate them in a closed environment filled with ethylene mixed gas for 24-48 hours, and screen out orange or red colonies. Microorganisms include genetically engineered bacteria introduced with exogenous MVA and / or MEP metabolic pathways, and any of the microorganisms naturally containing the above metabolic pathways. The mixed gas includes ethylene and air. This method is low in cost, easy to operate, and has obvious color development, and has the advantages of high screening throughput and short cycle. The above screening method is used to optimize the B. trispora strains producing β-carotene or lycopene, which can make the strains develop color in a short time, which is beneficial to the strains with high-throughput carotenoid metabolic flux. Identification, efficient and convenient.

The present invention discloses an apparatus for forming and screening a large-scale single-layer two-dimensional liquid droplet array, and a use method thereof. The apparatus comprises a micro-column array chip, a scraping plate for auxiliary spreading of a liquid droplet, a probe for controlling the liquid droplet, a three-dimensional translation table and other components, wherein a micro groove unit for trapping the single liquid droplet is formed by using the gap between micro-columns, and the liquid droplet is spread in the micro groove unit in a single-layer and single manner through gravity effect and capillary effect or through an auxiliary means so as to form the single-layer liquid droplet array. With the apparatus of the present invention, the accurate positioning and detection on the liquid droplet, the sucking on the target liquid droplet and the subsequent treatments can be performed. According to the present invention, the apparatus has characteristics of simple structure and easy operation; and the method has advantages of fast liquid droplet spreading, high flux, high liquid droplet trapping efficiency, liquid droplet position fixing and the like, and is suitable for single cell analysis and screening, single molecule analysis and screening, high-throughput gene screening, protein directed evolution, antibody screening, microbial research, drug screening, and other fields.

The invention discloses a multi-channel droplet quantitative measuring device based on pressure driving. The multi-channel droplet quantitative measuring device comprises a sample application probe for performing sample application operation on a sample solution, a working medium liquid storage tank for containing a working medium, a pressure control system for controlling the pressure in the working medium liquid storage tank, and a fluid damper provided with a micro-channel structure and having a damping effect on fluid in the micro-channel structure, wherein the liquid inlet end of the fluid damper communicates with the medium liquid storage tank, and the liquid outlet end of the fluid damper communicates with the liquid inlet end of the sample application probe in a sealed mode. The liquid droplet quantitative measuring device has the liquid droplet quantitative measuring capacity from picoliter level to microliter level and the liquid droplet quantitative measuring precision as low as picoliter level. According to the multi-channel droplet quantitative measuring device, various solutions with different viscosities can be measured in parallel, and the capability is an essentialcapability for large-scale high-throughput screening applications such as drug screening, protein crystallization condition screening and the like which need to treat a large amount of solutions withdifferent properties. The device is easy to construct, convenient to operate and good in reliability.